CN114591967A

CN114591967A - Application of corn TCP gene in cross breeding

Info

Publication number: CN114591967A
Application number: CN202210204361.2A
Authority: CN
Inventors: 刘宇婷; 戴周燕; 王海洋; 王胡海灵; 杨盼盼; 吴骏滔
Original assignee: South China Agricultural University
Current assignee: South China Agricultural University
Priority date: 2022-03-02
Filing date: 2022-03-02
Publication date: 2022-06-07
Anticipated expiration: 2042-03-02
Also published as: CN114591967B

Abstract

The invention discloses application of two corn TCP genes in cross breeding. The invention successfully carries out site-directed editing on the ZmTCP11 and the ZmTCP15 genes by using a CRISPR/Cas9 gene editing technology to obtain a plurality of homozygous strains mutated at target sites. By comparing the phenotype with the wild type and the single mutant strain thereof, the fact that the plant only shows the female structure phenotype of the tassel due to the double-gene function deletion is found, the plant continues to develop to form additional grains in the later period, other basic agronomic characters are not influenced, and the final yield cannot be reduced. The ZmTCP11 and ZmTCP15 double-gene function deletion mutant can save the step of female parent material breeding in the cross breeding process, the step of castration before cross breeding is changed into the step of bagging the male ear, the homozygous female parent seed can be recovered, the production efficiency is improved, and in addition, the double-gene function deletion mutant is also an excellent mutant for researching the male ear sex differentiation molecule regulation and control mechanism of corn.

Description

Application of corn TCP gene in cross breeding

Technical Field

The invention relates to the technical field of corn crossbreeding, in particular to application of two corn TCP genes ZmTCP11 and ZmTCP15 in corn crossbreeding.

Background

Corn (Latin's name: Zea mays L.) is an annual herbaceous plant of Zea of Gramineae, corn is an annual cross-pollinated plant with the same plants, namely male and female, is tall and strong in stem, is an important food crop and feed crop, is also a crop with the highest total yield all over the world, and has the planting area and the total yield only second to rice and wheat.

The crossbreeding is a breeding method in which different varieties are used as male and female parents for hybridization to form different genetic diversity combinations, and then a new variety with excellent male and female characteristics is obtained by screening, and the crossbreeding method is widely applied to grain crops such as rice, corn and the like. The corn is a male and female isogenous cross-flowering plant, and cross-pollination enables cross breeding to be simple, feasible and economical in the corn, and the cross breeding is widely applied to corn production.

The positions of the male ear inflorescence and the female ear inflorescence of the corn are different, the male ear at the top belongs to a cone inflorescence structure with limited branches, the female ear is positioned at the axilla of the corn stalk, is a scion inflorescence structure with thickened scion stalk, and is wrapped in bracts on the short branches of the scion. In the cross breeding process, the pollen of the male ear of the female parent is often removed before the pollen of the male ear of the female parent is scattered, so that the cross pollination process is avoided being influenced, and meanwhile, the female parent material used in the breeding process needs to be regularly bred and can be continuously used, so that the land resource and the labor resource cost in the cross breeding process are increased.

The TCP family is used as a transcription factor specific to a plant and is widely involved in the growth and development regulation of various plant groups at multiple stages. Wherein the Class II TCP transcription factor subfamily plays a core regulation function in the leaf development, the lateral branch formation and the flower development of plants. ZmTCP11 and ZmTCP15 are two TCP Class II genes directly homologous with a sorghum yield increasing gene MSD1 in corn, and the effects of ZmTCP11 and ZmTCP15 on the molecular regulation of corn tassel sex differentiation are not reported at present.

Disclosure of Invention

The invention aims to overcome the defect of complicated breeding of female parent materials in the prior crossbreeding technology, and provides the application of ZmTCP11 and ZmTCP15 genes and/or double mutations thereof in regulating and controlling the sex determination of maize tassels or culturing maize varieties with tassels being feminized, thereby being used for breeding of the crossbreeding female parent materials.

The second purpose of the invention is to provide the application of ZmTCP11 and ZmTCP15 genes and/or double mutation thereof in researching the sex differentiation of maize tassels.

The above object of the present invention is achieved by the following technical solutions:

according to the invention, after two TCP Class II genes ZmTCP11 and ZmTCP15 which are directly homologous with a sorghum yield increasing gene MSD1 in corn Kn5585 are knocked out by using a CRISPR/Cas9 transgenic editing technology, the observation shows that a double-gene function deletion mutant plant shows a tassel female fructification phenotype, but other basic agronomic traits are not influenced. And single gene loss of function mutants of both ZmTCP11 and ZmTCP15 did not exhibit an outward phenotype that is different from wild-type (WT). Through observation and statistics of basic agronomic traits of single-gene and double-gene function deletion mutants of ZmTCP11 and ZmTCP15 in field experiments, results show that the basic agronomic traits and wild types of the single-gene and double-gene function deletion mutants of ZmTCP11 and ZmTCP15 are not significantly different, and show that the genes of ZmTCP11 and ZmTCP15 play an important role in regulating and controlling the determination of maize tassel sex and have little influence on maize yield. Therefore, the obtained ZmTCP11 and ZmTCP15 double-gene function deletion mutant can have more remarkable advantages in the preparation of corn hybrid, the double mutant is used as a female parent for hybrid breeding, on one hand, the male ear is bagged in the early development stage of the male ear, so that female parent progeny can be obtained on the male ear, and the seed purity is efficiently ensured; on the other hand, the ear character is not affected, and the yield of normal hybrid seed production is not reduced. By applying the mutant, the female parent does not need to be bred additionally, the cost such as land and human resources can be effectively saved, and the production efficiency is improved. Meanwhile, the double mutants are excellent mutants for researching the corn tassel sex differentiation molecule regulation and control mechanism, and provide an important theoretical basis for corn molecule genetic improvement breeding. In conclusion, the invention has great theoretical and application prospects for corn crossbreeding and molecular genetic improvement breeding.

The invention therefore claims the following new uses for the ZmTCP11 and ZmTCP15 genes:

the use of the genes ZmTCP11 and ZmTCP15 and/or double mutations thereof in the regulation of maize tassel sex determination.

Specifically, the regulation of the maize tassel sex determination is the control of maize tassel estrification.

The ZmTCP11 and ZmTCP15 gene and/or its double mutation are used in breeding corn variety with male ear gynogenesis and fructification phenotype; and further using the strain in breeding of a hybridization breeding female parent material.

Specifically, the breeding application of the hybrid breeding female parent material is that the homozygous female parent material is prepared from the tassels of the hybrid female parent by utilizing the double-mutant tasseling fructification phenotype of the ZmTCP11 and ZmTCP15 genes, and the tassels are used for hybridization without influencing the yield of the hybrid.

The ZmTCP11 and ZmTCP15 gene and/or their double mutation are used in researching corn tassel sex differentiation.

Specifically, the nucleotide sequence of the ZmTCP11 gene is shown as SEQ ID NO. 1, and the nucleotide sequence of the ZmTCP15 gene is shown as SEQ ID NO. 2.

Specifically, the double mutation is a ZmTCP11 and ZmTCP15 double gene function deletion mutation.

Specifically, the ZmTCP11 and ZmTCP15 genes and/or double mutations thereof are knock-out ZmTCP11 and ZmTCP15 genes in corn to obtain ZmTCP11 and ZmTCP15 double-mutation transgenic corn plants.

Preferably, the ZmTCP11 and ZmTCP15 genes in maize are knocked out using CRISPR/Cas9 transgene editing technology.

Preferably, the sgRNA sequence of the ZmTCP11 gene is shown in SEQ ID NO 3-4, and the sgRNA sequence of the ZmTCP15 gene is shown in SEQ ID NO 5-6.

Further preferably, in the ZmTCP11 and ZmTCP15 double-mutation transgenic maize plant, the nucleotide sequence of the mutated ZmTCP11 is shown as SEQ ID NO. 7, and the nucleotide sequence of the mutated ZmTCP15 is shown as SEQ ID NO. 9.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides applications of ZmTCP11 and ZmTCP15 genes and/or double mutations thereof in regulating and controlling maize tassel sex determination or breeding maize varieties with tassel female. According to the invention, after corn ZmTCP11 and ZmTCP15 are knocked out by using a CRISPR/Cas9 transgenic editing technology, the observation shows that the double-gene function deletion mutant plant shows a male ear gynogenation fruiting phenotype, but other basic agronomic characters are not influenced, so that the obtained ZmTCP11 and ZmTCP15 double-gene function deletion mutant can be used as a female parent in hybrid breeding, the male ear is used for preparing a female parent homozygous material, and the female ear is used for hybrid seed production without influencing yield. And the single gene function deletion mutant of the ZmTCP11 and the ZmTCP15 does not show the appearance phenotype different from the Wild Type (WT), so the double gene function deletion mutant is also an excellent mutant for researching the corn tassel sex differentiation molecule control mechanism, and can better concentrate on the research of the tassel floret sex evolution process.

Drawings

FIG. 1 is a phylogenetic tree of the maize TCP Class II gene.

FIG. 2 is the genotyping of ZmTCP11 and ZmTCP15 transgenic plants. Mutant gene structure, blue represents the knockout target site, red represents the mutation site.

FIG. 3 shows the tcp11-2, tcp15-4, tcp15-8 and tcp11-1tcp15-4 mutant phenotypes. Pictures of whole corn (scale: 30cm), tassel and ear (scale: 5 cm).

FIG. 4 shows the tassel phenotype of the tcp11-1tcp15-4 double mutant.

FIG. 5 shows statistics of basic agronomic traits of WT, tcp11-2, tcp15-4, tcp15-8 and tcp11-1tcp 15-4. Plant height, ear height, tassel branch number, ear length, ear width, ear row number, row grain number, and hundred grain weight.

FIG. 6 is an observation of tassel florets from wild type maize and the tcp11-1tcp15-4 mutant. a. Wild type V7 early tassels florets; b. the late stage of wild type V8 tassel and spikelet; c. tassel spikelet at wild type V11 stage; d. the male ear is superior in the wild type V11 period; e. the male ear is superior in the wild type V11 period; f. maize tcp11-1tcp15-4 mutant V7 early tassels; g. maize tcp11-1tcp15-4 mutant V8 late tassel superior flowers; h. tassels in maize stage tcp11-1tcp15-4 mutant V11; i. maize tappets superior flowers at stage tcp11-1tcp15-4 mutant V11; j. maize under tassel flowers at time stage tcp11-1tcp15-4 mutant V11. Wherein, the scale of a and f is 200 μm, the scale of b and g is 0.5mm, and the scale of c, d, e, h, i and j is 1 mm.

Detailed Description

The invention is further described with reference to the drawings and specific examples, which are not intended to limit the invention in any way. Reagents, methods and apparatus used in the present invention are conventional in the art unless otherwise indicated.

Unless otherwise indicated, reagents and materials used in the following examples are commercially available.

Example 1 construction and genotyping of CRISPR/Cas9 transgenic plants of ZmTCP11 and ZmTCP15 genes

We firstly analyze the corn TCP Class II gene, and add part of the reported Arabidopsis thaliana, rice and sorghum TCP Class II gene to construct phylogenetic evolution analysis (figure 1), the results show that two TCP Class II genes ZmTCP11 and ZmTCP15 in corn are directly homologous with the sorghum yield increasing gene MSD1, the nucleotide sequence of the ZmTCP11 gene is shown as SEQ ID NO:1, and the nucleotide sequence of the ZmTCP15 gene is shown as SEQ ID NO: 2.

In order to obtain functional deletion mutants of ZmTCP11 and ZmTCP15, the structures of the two genes are analyzed, two sgRNA sequences are respectively selected on ZmTCP11 and ZmTCP15 according to the standard of designing 5'-G- (N)19-NGG-3' by a CRISPR/Cas9 gene knockout system target spot, and the sequences are shown as SEQ ID NO: 3-6:

ZmTCP11 sgRNA：CCACAGCAAGATCCGCACGGCGC(SEQ ID NO:3)；

GACGGACTCGAGGGAGGAGCTGG(SEQ ID NO:4)；

ZmTCP15 sgRNA：CCACAGCAAGATCCGCACGGCGC(SEQ ID NO:5)；

GCCGACGACCGACGGTGGTGAGG(SEQ ID NO:6)；

constructing transgenic knockout vectors of ZmTCP11 and ZmTCP15 by using a genetic engineering means, adopting an agrobacterium-mediated method, taking Basta as screening resistance, using a KN5585 self-bred line as a transformation background material to prepare a transgenic material, screening T0-generation positive seedlings by the Basta resistance, and recovering T1-generation corn transgenic seeds.

Through the identification and the separation of transgenic plants, single and double gene function deletion mutants of ZmTCP11 and ZmTCP15 are found, and are ZmTCP11 single mutants respectively: tcp11-2, ZmTCP15 single mutant: tcp15-4, tcp15-8 and double mutants of ZmTCP11 and ZmTCP 15: tcp11-1tcp 15-4; the specific gene mutation type is shown in figure 2, wherein the gene mutation type sequence is SEQ ID NO. 7: tcp 11-1; 8, SEQ ID NO: tcp 11-2; 9 of SEQ ID NO: tcp 15-4; 10, SEQ ID NO: tcp 15-8.

Example 2 phenotypic Observation of Single and Dual Gene loss of function mutants of ZmTCP11 and ZmTCP15

Wild corn and four isolated single-double gene function deletion mutants of ZmTCP11 and ZmTCP15 were planted in transgenic test fields of Hainan Mitsui and Hebei gallery: tcp11-2, tcp15-4, tcp15-8 and tcp11-1tcp15-4, the basic agronomic traits of the mutant material and wild type corn were observed by field experiments and statistically correlated data.

It was observed that the sexual differentiation of some florets on tassels of the maize tcp11-1tcp15-4 double gene loss mutant was impaired, hermaphrodite double sexual florets were produced and extended out of the slender style and could set (FIG. 4). The tcp11-1tcp15-4 mutant has partial floret sexual differentiation disorder except for the tassel, and other basic agronomic traits do not show significant difference with wild corn; during subsequent development, the gynoecial tassel continues to develop to form additional kernels, resulting in a ear phenotype. The foreign traits of the other three single-gene loss-of-function mutants, tcp11-2, tcp15-4 and tcp15-8, were also not significantly different from wild-type maize (FIG. 3). Statistics also showed that these mutant materials had no significant differences from the basic agronomic traits (plant height, ear height, tassel branch number, ear length, ear width, ear row number, row grain number and hundred grain weight) of wild type corn (fig. 5). The results show that ZmTCP11 and ZmTCP15 have the function of specifically regulating and controlling the sex determination of the maize tassel, and the double-mutant tassels thereof are female and fruited without affecting the yield of the tassels.

Example 3 Observation of tassel florets from Wild Type (WT) maize and the tcp11 tcp15 mutant

Tassel spikelets of wild-type maize and tcp11-1tcp15-4 mutants with two stages of seven mature leaves (V7) and eleven mature leaves (V11) were selected and observed using scanning and stereomicroscope.

The results show that there was no significant difference in the development of primordia between tassel florets of wild type maize at stage V7 (FIG. 6a) and of the tcp11-1tcp15-4 mutant (FIG. 6 f); however, by the time of V8, the carpel primordium in the wild-type maize tassel florets began to degenerate and abortive (FIG. 6b), while the carpel primordium of the superior flower in the tcp11-1tcp15-4 mutant tassel florets was retained and continued to develop in bulk (FIG. 6 g); by the time of V11, all florets on the wild-type maize tassel developed into unisexual tassels (FIGS. 6c, d, e), while the superior flowers in the tassel spikelet of the tcp11-1tcp15-4 mutant developed feminized carpel structures and extended long and thin style pillars (FIGS. 6h, i, g). It was shown that the tassel florets of the tcp11-1tcp15-4 mutant differentiated from wild-type maize during the differentiation period of V8-V10 sex.

Therefore, the obtained ZmTCP11 and ZmTCP15 double-gene function deletion mutant can have more remarkable advantages in the preparation of corn hybrid, the double mutant is used as a female parent for hybrid breeding, on one hand, the male ear is bagged in the early development stage of the male ear, so that female parent progeny can be obtained on the male ear, and the seed purity is efficiently ensured; on the other hand, the property of the female ear is not affected, and the yield of normal hybrid seed production is not reduced. The double mutants are excellent mutants for researching the corn tassel sex differentiation molecular regulation and control mechanism, and provide an important theoretical basis for corn molecular genetic improvement breeding.

Sequence listing

<110> southern China university of agriculture

Application of <120> corn TCP gene in cross breeding

<141> 2022-03-02

<160> 10

<170> SIPOSequenceListing 1.0

<210> 1

<211> 723

<212> DNA

<213> corn (Zea mays L.)

<400> 1

atgccgtcga ccgcgatgtc ttgggacggg tacggcgcgc agatcttcct cgccgacatg 60

tcgttccacc accaggatac cctggaggcg gtgttccagc agcctgcaga gacgacgctc 120

ctgcaggcgc cggcggggga gaaggagctg cccagcaacg ggtcgccgcc ggtggtggat 180

gacgccggcg tccaggccgc ggccgtacta cggaagcggc cgttccggac ggatcgccac 240

agcaagatcc gcacggcgca gggcgtccgc gaccgccgga tgcggctctc ggtcggggtc 300

gcgcgagagt tcttcgcgct gcaggaccgc ctcgggttcg acaaggccag caagaccgtg 360

aactggctcc tcgcccagtc caagccggcc atcgaccgcc tcgtcgacgc cgccgaccag 420

gcggctgtag tcacaggagg aggaccgacg ctggtgaagg ggagagggga ggggagcaac 480

tcaagcactt gctgtttgac ggactcgagg gaggagctgg ccacggagaa gggaagaagc 540

agagccggtg acgccggtct tgattggcca ccgcctctta ttgaagaaca cggctgcggc 600

gagctgggat ggatcatgtc ggaggcaaca gcaacgccgg agcaggacgg gctggagtgc 660

tactaccagt attgcctgca gctcgaggag ttgatgagat gcaacggcgg aatgccaagg 720

tga 723

<210> 2

<211> 825

<212> DNA

<213> corn (Zea mays L.)

<400> 2

atgccctcga ccgccgcgat gtcgtcgtct tgggacgggt acggcgggca gatcttcccc 60

gcagacatgt cgtcgttcca ccgccaccag gacaccctgg aggcggtgtt ccaccacccg 120

ccgcctgaga cgacgacgac gacggggctg ctgaggaacg acgggtcatc accggtggtg 180

gacgacggcg gcggccacgc cgcaccgcga aggcggccgt tccggacgga ccgccacagc 240

aagatccgca cggcgcaggg cgtgcgcgac cgccggatgc ggctgtcggt cggggtcgcg 300

cgcgagttct tcgcgctgca ggaccgcctc ggcttcgaca aggccagcaa gacggtgaac 360

tggctcctca cccagtccaa gccggccatc gaccgcctcg tcgacgccgc cgccgccgcc 420

gacgaccccg cggccgtagc agcctcagga ggccgacgac cgacggtggt gaggggcaga 480

ggcgagggca gctcctcgag cacttgctgc tgcttgacgg actcgagaga ggccgccgag 540

gaggcgacgg ggaacgggag aagcagaggc ggccctgacg acgggccacc ggcagcgctt 600

ctggaaggac acggcggctg cggcgagctg ggctggatca tgtcgggagc gcccacagca 660

gcggtggcaa cgacgacgac gacgacgccg cagcagccgg acgggcacga gtactactac 720

cagtattgcc tgcagctcga ggagatgatg cgatgcagca acgacgaagg agaaacaacg 780

ccaggtgatt tcttgtatgg tatgcagacg cgtgataggt cttga 825

<210> 3

<211> 23

<212> DNA

<213> corn (Zea mays L.)

<400> 3

ccacagcaag atccgcacgg cgc 23

<210> 4

<211> 23

<212> DNA

<213> corn (Zea mays L.)

<400> 4

gacggactcg agggaggagc tgg 23

<210> 5

<211> 23

<212> DNA

<213> corn (Zea mays L.)

<400> 5

ccacagcaag atccgcacgg cgc 23

<210> 6

<211> 23

<212> DNA

<213> corn (Zea mays L.)

<400> 6

gccgacgacc gacggtggtg agg 23

<210> 7

<211> 451

<212> DNA

<213> corn (Zea mays L.)

<400> 7

atgccgtcga ccgcgatgtc ttgggacggg tacggcgcgc agatcttcct cgccgacatg 60

tcgttccacc accaggatac cctggaggcg gtgttccagc agcctgcaga gacgacgctc 120

ctgcaggcgc cggcggggga gaaggagctg cccagcaacg ggtcgccgcc ggtggtggat 180

gacgccggcg tccaggccgc ggccgtacta cggaagcggc cgttccggac ggatcgccac 240

agagctggcc acggagaagg gaagaagcag agccggtgac gccggtcttg attggccacc 300

gcctcttatt gaagaacacg gctgcggcga gctgggatgg atcatgtcgg aggcaacagc 360

aacgccggag caggacgggc tggagtgcta ctaccagtat tgcctgcagc tcgaggagtt 420

gatgagatgc aacggcggaa tgccaaggtg a 451

<210> 8

<211> 549

<212> DNA

<213> corn (Zea mays L.)

<400> 8

atgccgtcga ccgcgatgtc ttgggacggg tacggcgcgc agatcttcct cgccgacatg 60

tcgttccacc accaggatac cctggaggcg gtgttccagc agcctgcaga gacgacgctc 120

ctgcaggcgc cggcggggga gaaggagctg cccagcaacg ggtcgccgcc ggtggtggat 180

gacgccggcg tccaggccgc ggccgtacta cggaagcggc cgttccggac ggatcgccac 240

agagccggcc atcgaccgcc tcgtcgacgc cgccgaccag gcggctgtag tcacaggagg 300

aggaccgacg ctggtgaagg ggagagggga ggggagcaac tcaagcactt gctgtttgac 360

ggactcgagg gaggtgacgc cggtcttgat tggccaccgc ctcttattga agaacacggc 420

tgcggcgagc tgggatggat catgtcggag gcaacagcaa cgccggagca ggacgggctg 480

gagtgctact accagtattg cctgcagctc gaggagttga tgagatgcaa cggcggaatg 540

ccaaggtga 549

<210> 9

<211> 684

<212> DNA

<213> corn (Zea mays L.)

<400> 9

atgccctcga ccgccgcgat gtcgtcgtct tgggacgggt acggcgggca gatcttcccc 60

gcagacatgt cgtcgttcca ccgccaccag gacaccctgg aggcggtgtt ccaccacccg 120

ccgcctgaga cgacgacgac gacggggctg ctgaggaacg acgggtcatc accggtggtg 180

gacgacggcg gcggccacgc cgcaccgcga aggcggccgt tccggacgga ccgccacaga 240

gccggccatc gaccgcctcg tcgacgccgc cgccgacgcc gacgaccccg cggcagtagc 300

agcctcagga ggccgacgac cgacggtgtg aggggcagag gcgagggcag ctcctcgagc 360

acttgctgct gcttgacgga ctcgagagag gccgccgagg aggcgacggg gaacgggaga 420

agcagaggcg gccctgacga cgggccaccg gcagcgcttc tggaaggaca cggcggctgc 480

ggcgagctgg gctggatcat gtcgggagcg cccacagcag cggtggcaac gacgacgacg 540

acgacgccgc agcagccgga cgggcacgag tactactacc agtattgcct gcagctcgag 600

gagatgatgc gatgcagcaa cgacgaagga gaaacaacgc caggtgattt cttgtatggt 660

atgcagacgc gtgataggtc ttga 684

<210> 10

<211> 745

<212> DNA

<213> corn (Zea mays L.)

<400> 10

atgccctcga ccgccgcgat gtcgtcgtct tgggacgggt acggcgggca gatcttcccc 60

gcagacatgt cgtcgttcca ccgccaccag gacaccctgg aggcggtgtt ccaccacccg 120

ccgcctgaga cgacgacgac gacggggctg ctgaggaacg acgggtcatc accggtggtg 180

gacgacggcg gcggccacgc cgcaccgcga aggcggccgt tccggacggc gcagggcgtg 240

cgcgaccgcc tcggcttcga caaggccagc aagacggtga actggctcct cacccagtcc 300

agccggccat cgaccgcctc gtcgacgccg ccgccgccgc cgacgacccc gcggccgtag 360

cagcctcagg aggccgacga ccgacggtgt gaggggcaga ggcgagggca gctcctcgag 420

cacttgctgc tgcttgacgg actcgagaga ggccgccgag gaggcgacgg ggaacgggag 480

aagcagaggc ggccctgacg acgggccacc ggcagcgctt ctggaaggac acggcggctg 540

cggcgagctg ggctggatca tgtcgggagc gcccacagca gcggtggcaa cgacgacgac 600

gacgacgccg cagcagccgg acgggcacga gtactactac cagtattgcc tgcagctcga 660

ggagatgatg cgatgcagca acgacgaagg agaaacaacg ccaggtgatt tcttgtatgg 720

tatgcagacg cgtgataggt cttga 745

Claims

Use of the genes ZmTCP11 and ZmTCP15 for regulating maize tassel gender determination.
Use of double mutations in the ZmTCP11 and ZmTCP15 genes in the regulation of maize tassel gender determination.
3. The use of claim 1 or 2, wherein the modulation of maize tassel sex determination is the control of maize tassel gynogenesis and fructification.
Use of the genes ZmTCP11 and ZmTCP15 for breeding maize varieties with tassel gynogenesis and fruiting phenotype.
The application of the double mutation of ZmTCP11 and ZmTCP15 genes in breeding corn variety with tassel female fructification phenotype.
6. The use of any one of claims 1 to 5, wherein the nucleotide sequence of the ZmTCP11 gene is shown as SEQ ID NO. 1, and the nucleotide sequence of the ZmTCP15 gene is shown as SEQ ID NO. 2.
7. The use of claim 2 or 5, wherein the double mutation is a deletion mutation of the ZmTCP11 and ZmTCP15 double gene function.
8. The use of any one of claims 3 to 5, wherein ZmTCP11 and ZmTCP15 genes in maize are knocked out to obtain ZmTCP11 and ZmTCP15 double-mutation transgenic maize plants.
9. The use according to claim 8, wherein the ZmTCP11 and ZmTCP15 genes in maize are knocked out by using CRISPR/Cas9 transgene editing technology.
10. The application of claim 9, wherein the sgRNA sequence of the ZmTCP11 gene is shown as SEQ ID NO 3-4, and the sgRNA sequence of the ZmTCP15 gene is shown as SEQ ID NO 5-6.